Allocyclic behaviour and underreplication of the nucleolus chromosome in Pseudodiamesa (Chironomidae)

The chironomid midge Pseudodiamesa branickii shows three polytene, banded elements in salivary gland and Malpighian tubule cells, none of which carry a nucleolar organizer. The present investigation revealed four chromosome pairs in mitotic metaphases, the nucleolus was localized in the tiny chromosome 4. Microphotometric determinations of the DNA contents in mitotic and salivary gland nuclei of Ps. branickii yielded a 2C value of 0.23 pg DNA and a maximum level of 12 endoreplications. In both salivary glands and Malpighian tubules, chromosome 4 appeared as a nonbanded network of chromatin in Feulgen preparations. Indirect immunofluorescent staining with antibodies specific for RNA X DNA hybrids indicated that this structure was transcriptionally active. Its reaction to heavy metal staining suggested that it included the nucleolar organizer, and this was conclusively demonstrated by in situ hybridization with 125I-rRNA. Chromosome pair 4 comprises about 11.4% of the total DNA in metaphase, but only 3.7% in the highly polytene salivary gland complement. According to a numerical simulation, the degree of underrepresentation suggests that about 70% of chromosome 4 is precluded from polytenization. The allocyclic aspect is probably due to this underreplication, with the nucleolar activity causing a structural loosening of the rest of the chromosome. The possible nature of the underreplicated sequences is discussed in the light of recent work on molecular aspects of rDNA structure in various Diptera.